Light-sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development.

Background: Selective plane illumination microscopy (SPIM a type of light-sheet microscopy) involves focusing a thin sheet of laser light through a specimen at right angles to the objective lens. As only the thin section of the specimen at the focal plane of the lens is illuminated, out of focus light is naturally absent and toxicity due to light (phototoxicity) is greatly reduced enabling longer term live imaging. OpenSPIM is an open access platform (Pitrone et al. 2013 and OpenSPIM.org) created to give new users step-by-step instructions on building a basic configuration of a SPIM microscope, which can in principle be adapted and upgraded to each laboratory’s own requirements and budget. Here we describe our own experience with the process of designing, building, configuring and using an OpenSPIM for our research into the early development of the polyclad flatworm Maritigrella crozieri – a non-model animal. Results: Our OpenSPIM builds on the standard design with the addition of two colour laser illumination for simultaneous detection of two probes/molecules and dual sided illumination, which provides more even signal intensity across a specimen. Our OpenSPIM provides high resolution 3d images and time lapse recordings, and we demonstrate the use of two colour lasers and the benefits of two color dual-sided imaging. We used our microscope to study the development of the embryo of the polyclad flatworm M. crozieri. The capabilities of our microscope are demonstrated by our ability to record the stereotypical spiral cleavage pattern of M. crozieri with high-speed multi-view time lapse imaging. 3D and 4D (3D + time) reconstruction of early development from these data is possible using image registration and deconvolution tools provided as part of the open source Fiji platform. We discuss our findings on the pros and cons of a self built microscope. Conclusions: We conclude that home-built microscopes, such as an OpenSPIM, together with the available open source software, such as MicroManager and Fiji, make SPIM accessible to anyone interested in having continuous access to their own light-sheet microscope. However, building an OpenSPIM is not without challenges and an open access microscope is a worthwhile, if significant, investment of time and money. Multi-view 4D microscopy is more challenging than we had expected. We hope that our experience gained during this project will help future OpenSPIM users with similar ambitions

Asteroseismology of the Hyades with K2: first detection of main-sequence solar-like oscillations in an open cluster

The Hyades open cluster was targeted during Campaign 4 (C4) of the NASA K2 mission, and short-cadence data were collected on a number of cool main-sequence stars. Here, we report results on two F-type stars that show detectable oscillations of a quality that allows asteroseismic analyses to be performed. These are the first ever detections of solar-like oscillations in main-sequence stars in an open cluster.Comment: 12 pages, 8 figures, 2 tables; accepted for publication in MNRA

Detection of solar-like oscillations in relics of the Milky Way: asteroseismology of K giants in M4 using data from the NASA K2 mission

Asteroseismic constraints on K giants make it possible to infer radii, masses and ages of tens of thousands of field stars. Tests against independent estimates of these properties are however scarce, especially in the metal-poor regime. Here, we report the detection of solar-like oscillations in 8 stars belonging to the red-giant branch and red-horizontal branch of the globular cluster M4. The detections were made in photometric observations from the K2 Mission during its Campaign 2. Making use of independent constraints on the distance, we estimate masses of the 8 stars by utilising different combinations of seismic and non-seismic inputs. When introducing a correction to the Delta nu scaling relation as suggested by stellar models, for RGB stars we find excellent agreement with the expected masses from isochrone fitting, and with a distance modulus derived using independent methods. The offset with respect to independent masses is lower, or comparable with, the uncertainties on the average RGB mass (4-10%, depending on the combination of constraints used). Our results lend confidence to asteroseismic masses in the metal poor regime. We note that a larger sample will be needed to allow more stringent tests to be made of systematic uncertainties in all the observables (both seismic and non-seismic), and to explore the properties of RHB stars, and of different populations in the cluster.Comment: 6 pages, 3 figures, accepted for publication in MNRA

Circulating exosomes deliver free fatty acids from the bloodstream to cardiac cells:Possible role of CD36

Regulation of circulating free fatty acid (FFA) levels and delivery is crucial to maintain tissue homeostasis. Exosomes are nanomembranous vesicles that are released from diverse cell types and mediate intercellular communication by delivering bioactive molecules. Here, we sought to investigate the uptake of FFAs by circulating exosomes, the delivery of FFA-loaded exosomes to cardiac cells and the possible role of the FFA transporter CD36 in these processes. Circulating exosomes were purified from the serum of healthy donors after an overnight fast (F) or 20 minutes after a high caloric breakfast (postprandial, PP). Western blotting, Immunogold Electron Microscopy and FACS analysis of circulating exosomes showed that CD36 was expressed under both states, but was higher in postprandial-derived exosomes. Flow cytometry analysis showed that circulating exosomes were able to take-up FFA directly from serum. Importantly, preincubation of exosomes with a blocking CD36 antibody significantly impeded uptake of the FFA analogue BODIPY, pointing to the role of CD36 in FFA exosomal uptake. Finally, we found that circulating exosomes could delivery FFA analogue BODIPY into cardiac cells ex vivo and in vivo in a mice model. Overall, our results suggest a novel mechanism in which circulating exosomes can delivery FFAs from the bloodstream to cardiac tissue. Further studies will be necessary to understand this mechanism and, in particular, its potential involvement in metabolic pathologies such as obesity, diabetes and atherosclerosis

Limits on surface gravities of Kepler planet-candidate host stars from non-detection of solar-like oscillations

We present a novel method for estimating lower-limit surface gravities log g of Kepler targets whose data do not allow the detection of solar-like oscillations. The method is tested using an ensemble of solar-type stars observed in the context of the Kepler Asteroseismic Science Consortium. We then proceed to estimate lower-limit log g for a cohort of Kepler solar-type planet-candidate host stars with no detected oscillations. Limits on fundamental stellar properties, as provided by this work, are likely to be useful in the characterization of the corresponding candidate planetary systems. Furthermore, an important byproduct of the current work is the confirmation that amplitudes of solar-like oscillations are suppressed in stars with increased levels of surface magnetic activity.Comment: Accepted for publication in ApJ; 35 pages, 10 figures, 5 table

Characterizing two solar-type Kepler subgiants with asteroseismology: KIC10920273 and KIC11395018

Determining fundamental properties of stars through stellar modeling has improved substantially due to recent advances in asteroseismology. Thanks to the unprecedented data quality obtained by space missions, particularly CoRoT and Kepler, invaluable information is extracted from the high-precision stellar oscillation frequencies, which provide very strong constraints on possible stellar models for a given set of classical observations. In this work, we have characterized two relatively faint stars, KIC10920273 and KIC11395018, using oscillation data from Kepler photometry and atmospheric constraints from ground-based spectroscopy. Both stars have very similar atmospheric properties; however, using the individual frequencies extracted from the Kepler data, we have determined quite distinct global properties, with increased precision compared to that of earlier results. We found that both stars have left the main sequence and characterized them as follows: KIC10920273 is a one-solar-mass star (M=1.00 +/- 0.04 M_sun), but much older than our Sun (t=7.12 +/- 0.47 Gyr), while KIC11395018 is significantly more massive than the Sun (M=1.27 +/- 0.04 M_sun) with an age close to that of the Sun (t=4.57 +/- 0.23 Gyr). We confirm that the high lithium abundance reported for these stars should not be considered to represent young ages, as we precisely determined them to be evolved subgiants. We discuss the use of surface lithium abundance, rotation and activity relations as potential age diagnostics.Comment: 12 pages, 3 figures, 5 tables. Accepted by Ap

Modelling the Autocovariance of the Power Spectrum of a Solar-Type Oscillator

Asteroseismology is able to conduct studies on the interiors of solar-type stars from the analysis of stellar acoustic spectra. However, such an analysis process often has to rely upon subjective choices made throughout. A recurring problem is to determine whether a signal in the acoustic spectrum originates from a radial or a dipolar oscillation mode. In order to overcome this problem, we present a procedure for modelling and fitting the autocovariance of the power spectrum which can be used to obtain global seismic parameters of solar-type stars, doing so in an automated fashion without the need to make subjective choices. From the set of retrievable global seismic parameters we emphasize the mean small frequency separation and, depending on the intrinsic characteristics of the power spectrum, the mean rotational frequency splitting. Since this procedure is automated, it can serve as a useful tool in the analysis of the more than one thousand solar-type stars expected to be observed as part of the Kepler Asteroseismic Investigation (KAI). We apply the aforementioned procedure to simulations of the Sun. Assuming different apparent magnitudes, we address the issues of how accurately and how precisely we can retrieve the several global seismic parameters were the Sun to be observed as part of the KAI.Comment: 10 pages, 8 figures, accepted for publication in MNRA

Oscillation mode frequencies of 61 main sequence and subgiant stars observed by Kepler

Solar-like oscillations have been observed by Kepler and CoRoT in several solar-type stars, thereby providing a way to probe the stars using asteroseismology. We provide the mode frequencies of the oscillations of various stars required to perform a comparison with those obtained from stellar modelling. We used a time series of nine months of data for each star. The 61 stars observed were categorised in three groups: simple, F-like and mixed-mode. The simple group includes stars for which the identification of the mode degree is obvious. The F-like group includes stars for which the identification of the degree is ambiguous. The mixed-mode group includes evolved stars for which the modes do not follow the asymptotic relation of low-degree frequencies. Following this categorisation, the power spectra of the 61 main sequence and subgiant stars were analysed using both maximum likelihood estimators and Bayesian estimators, providing individual mode characteristics such as frequencies, linewidths, and mode heights. We developed and describe a methodology for extracting a single set of mode frequencies from multiple sets derived by different methods and individual scientists. We report on how one can assess the quality of the fitted parameters using the likelihood ratio test and the posterior probabilities. We provide the mode frequencies of 61 stars (with their 1-sigma error bars), as well as their associated echelle diagrams.Comment: 83 pages, 17 figures, 61 tables, paper accepted by Astronomy and Astrophysic

Ages and fundamental properties of Kepler exoplanet host stars from asteroseismology

We present a study of 33 {\it Kepler} planet-candidate host stars for which asteroseismic observations have sufficiently high signal-to-noise ratio to allow extraction of individual pulsation frequencies. We implement a new Bayesian scheme that is flexible in its input to process individual oscillation frequencies, combinations of them, and average asteroseismic parameters, and derive robust fundamental properties for these targets. Applying this scheme to grids of evolutionary models yields stellar properties with median statistical uncertainties of 1.2\% (radius), 1.7\% (density), 3.3\% (mass), 4.4\% (distance), and 14\% (age), making this the exoplanet host-star sample with the most precise and uniformly determined fundamental parameters to date. We assess the systematics from changes in the solar abundances and mixing-length parameter, showing that they are smaller than the statistical errors. We also determine the stellar properties with three other fitting algorithms and explore the systematics arising from using different evolution and pulsation codes, resulting in 1\% in density and radius, and 2\% and 7\% in mass and age, respectively. We confirm previous findings of the initial helium abundance being a source of systematics comparable to our statistical uncertainties, and discuss future prospects for constraining this parameter by combining asteroseismology and data from space missions. Finally we compare our derived properties with those obtained using the global average asteroseismic observables along with effective temperature and metallicity, finding an excellent level of agreement. Owing to selection effects, our results show that the majority of the high signal-to-noise ratio asteroseismic {\it Kepler} host stars are older than the Sun.Comment: 25 pages, 17 figures, MNRAS accepte